Conformational Epitope Mapping of C-Reactive Protein in Solution by Hydrogen/Deuterium Exchange Mass Spectrometry
10.19756/j.issn.0253-3820.241209
- VernacularTitle:基于氢氘交换质谱技术的C反应蛋白在溶液中的构象表位映射
- Author:
Hao-Feng SUN
1
;
Jian-Yi LIU
;
Qi ZHANG
;
Hui JIAO
;
Min ZHOU
;
De-Wei SONG
Author Information
1. 南京理工大学化学与化工学院,南京 210094;中国计量科学研究院化学计量与分析科学研究所,北京 100029
- Keywords:
C-reactive protein;
Hydrogen/deuterium exchange;
Epitope mapping;
Conformational epitope;
Allosteric targets
- From:
Chinese Journal of Analytical Chemistry
2025;53(4):631-639
- CountryChina
- Language:Chinese
-
Abstract:
C-Reactive protein(CRP)is an important acute-phase response protein,which is widely used in the assessment of inflammation and cardiovascular disease risk,and acts as a pathogenic factor directly involved in the disease process of certain conditions.Therefore,developing immunosuppressants targeting CRP or investigating its pathogenic mechanisms is of significant importance.Most B-cell epitopes are conformational epitopes,and studying conformational epitopes is typically challenging.To date,no methods have been reported for mapping the conformational epitopes of CRP in solution.In this study,a rapid strategy was developed for studying conformational epitopes by combining hydrogen/deuterium exchange mass spectrometry(HDX-MS)with multiparametric prediction of B-cell epitopes and protein secondary structure analysis.This approach was successfully applied to the binding sites and allosteric targets of the 115 kDa full pentameric CRP and the clinically used monoclonal antibody(mAb)5A8.The results showed that the amino acid residues 84-103,138-146,and 165-173 together form the potential conformational epitopes for mAb 5A8 on CRP,while the amino acid residues 21-32 and 175-178 were identified as potential allosteric targets.The discovery of the mAb 5A8 binding sites and allosteric targets was crucial for improving clinical diagnostic capabilities.Experimental results demonstrated that this workflow allowed rapid conformational epitope mapping of CRP under near-physiological conditions,with advantages such as high speed,high sensitivity,and high throughput.
